Toll样受体4在人滋养细胞中的表达及免疫作用

Study on the expression and immunologic effect of Toll like receptor 4 in human trophocyte

目的:探讨Toll样受体4(Toll like receptor 4,TLR4)在滋养细胞中的表达及其免疫作用。方法:采用RT-PCR分别检测原代培养滋养细胞和经1mg/LLPS刺激12h后的滋养细胞中TLR4 mRNA表达水平;流式细胞计数分别检测LPS刺激的滋养细胞中TLR4抗体封闭组、非TLR4抗体封闭组和空白对照组细胞凋亡发生率;ELISA法分别检测LPS刺激的滋养细胞中TLR4抗体封闭组、非TLR4抗体封闭组和空白对照组细胞TNF-α分泌水平。结果:RT-PCR结果显示,经LPS刺激的滋养细胞较正常滋养细胞TLR4mRNA表达水平明显增强;在LPS刺激的滋养细胞中,流式细胞计数结果显示,TLR4抗体封闭组和非TLR4抗体封闭组细胞凋亡率存在明显差异。ELISA结果显示,TLR4抗体封闭组和非TLR4抗体封闭组细胞TNF-α分泌水平也有显著差异。结论:TLR4信号途径在LPS引起的人滋养细胞凋亡及TNF-α分泌等炎性过程中发挥着重要作用。

Objective：To investigate the expression and immunologic effect of TLR4 in human trophocyte.Methods：Expression levels of TLR4 mRNA in normal and LPS-stimulated human trophocyte（1 mg/L LPS, 12 h） were detected by RT-PCR respectively; Incident rates of apoptosis were measured by Flow cytometry （FCM） in two groups of LPS-stimulated human trophocyte , which were antibody-blocked or none antibody-blocked cells, and other group of control cell respectively; Level of TNF-α was measured by enzyme linked immunosorbent assay （ELISA） in cells as above FCM measured respectively.Results：RT-PCR results showed that expression level of TLR4 mRNA in LPS-stimulated human trophocyte was higher than that in normal cell, differences had statistical significance（P〈0.01）; FCM results showed that incident rate of apoptosis had statistical differences in antibody-blocked or none antibody-blocked cells of LPS-stimulated human trophocyte （P〈0.05）.ELISA results showed that the level of TNF-αalso had statistical differences in antibody-blocked or none antibody-blocked cells of LPS-stimulated human trophocyte （P〈0.05）.Conclusion：TLR4 plays an important role during the inflammatory course of LPS-induced apoptosis and secretion of TNF-αin human trophocyte.